Inactivation of MARCH5 and Drp1 blocks pressure-induced mitochondrial fragmentation.

<p>(A) Differentiated RGC5 cells transfected with expression constructs for MARCH5^YFP or MARCH5^H43W-YFP were exposed for 72 hours to 30 mmHg elevated pressure or left untreated as control. Mitochondrial morphology was assessed following cytochrome <i>c</i> staining. The bar graph represents three independent experiments (>200 cell counted/condition) with * marking p<0.05 and ** marking p<0.01 (Student's t-test). Error bars correspond to SEM. (B) RGC5 cells expressing MARCH5 or MARCH5^H43W and photoactivatable GFP (PAGFP) were exposed to 30 mmHg for three days or left untreated as control and mitochondrial interconnectivity was measured by PAGFP diffusion after photoactivation and compared to ambient pressure, MARCH5 expressing cells. Analyzed were 20 cells/condition with the error bars representing SEM and ** marking p<0.01 and n.s. marking p>0.05 (Student's t-test). (C) Differentiated RGC5 cells transfected with expression constructs for Drp1^YFP or Drp1^K38A–YFP were treated as described in A.